The zoo announced on social media that the 49-year-old ape was euthanised on Wednesday morning, saying she had suffered from several health issues that slowly incapacitated her.
Josephine was born in the wild in March 1967 and arrived at Zoo Miami in March 1983.
Harambe was the name of a dead Western Lowland Gorilla shot and killed at Cincinnati Zoo in May after grabbing and dragging a three-year-old boy who fell into his enclosure.
The incident became a viral sensation with thousands of memes created by online commenters, many of whom were protesting what they believed to be an “unlawful” killing.
Local media reported that Josephine gave birth to her first offspring, a male named Moja, in 1984, who was the first gorilla born at Zoo Miami.
Moja moved to Gladys Porter Zoo in Texas, where he fathered several gorillas, including Harambe, who gained worldwide attention last year when zookeepers at the Cincinnati Zoo shot him to protect a child that fell in the enclosure.
Zoo Miami’s only remaining gorilla, Fredrika, will be transferred to another zoo.